title: Hedgehog signaling is active in human prostate cancer stroma and regulates proliferation and differentiation of adjacent epithelium.
abstract: BACKGROUND
  Contribution of stromal Hedgehog (Hh) signaling is evident in the prostate gland in mice, but needs translation to human tissues if Hh therapeutics are to be used effectively. Our goal was to determine if primary human prostate fibroblasts contain cilia, and respond to prostate Hh signaling.
  
  
  METHODS
  Primary human prostate cancer-associated (CAFs), and adjacent non-malignant (NPFs) fibroblasts isolated from human tissue specimens were analyzed using immunofluorescence, real-time PCR, and available array data. Cell culture and tissue recombination were used to determine responsiveness of human fibroblasts to Hh pathway manipulation and the paracrine effects of stromal Hh signaling, respectively.
  
  
  RESULTS
  Prostatic fibroblasts were capable of forming primary cilia, with the capacity for active Hh signaling as seen by Smo co-localization to the tip of the primary cilium. Expression of genes known to represent a signature of active Hh signaling in the prostate (especially Fgf5 and Igfbp6) were increased in CAFs compared to NPFs. The level of canonical Hh genes and prostate Hh signature genes were rarely synchronous; with lower doses of Purmorphamine/BMS-833923 regulating canonical transcription factors, and higher doses effecting prostate Hh signature genes. Grafts consisting of NPFs with constitutively active Hh signaling induced increased proliferation and dedifferentiation of adjacent non-malignant BPH-1 epithelial cells.
  
  
  CONCLUSIONS
  These data show that human prostatic fibroblasts have the capacity for Hh signaling and manipulation. Increased expression of a signature of prostatic Hh genes in the prostate tumor microenvironment suggests a role in the epithelial transformations driving prostate cancer (PCa).
authors.affiliation: Prostate Cancer Research Group, Department of Anatomy and Developmental Biology, Monash University, Clayton, Victoria, Australia; Department of Physiology, Monash University, Clayton, Victoria, Australia.
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Wilkinson
authors.forename: Luc
authors.initials: L
authors.lastname: Furic
authors.forename: Grant
authors.initials: G
authors.lastname: Buchanan
authors.forename: Ola
authors.initials: O
authors.lastname: Larsson
authors.forename: John
authors.initials: J
authors.lastname: Pedersen
authors.forename: Mark
authors.initials: M
authors.lastname: Frydenberg
authors.forename: Gail P
authors.initials: GP
authors.lastname: Risbridger
authors.forename: Renea A
authors.initials: RA
authors.lastname: Taylor
mesh_terms: D000818:Animals; D001549:Benzamides / Q000494:pharmacology; D002450:Cell Communication / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology*; D049109:Cell Proliferation; D002471:Cell Transformation, Neoplastic / Q000473:pathology; D002478:Cells, Cultured; D004848:Epithelium / Q000473:pathology; D005347:Fibroblasts / Q000473:pathology; D053823:Hedgehog Proteins / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D064593:Heterografts; D006801:Humans; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009025:Morpholines / Q000494:pharmacology; D011471:Prostatic Neoplasms / Q000473:pathology* / Q000503:physiopathology*; D011687:Purines / Q000494:pharmacology; D011799:Quinazolines / Q000494:pharmacology; D015398:Signal Transduction / Q000502:physiology*; D017154:Stromal Cells / Q000473:pathology
chemical_list: C583051:BMS-833923; D001549:Benzamides; D053823:Hedgehog Proteins; D009025:Morpholines; D011687:Purines; D011799:Quinazolines; C470893:purmorphamine
keywords: Hedgehog; microenvironment; paracrine signaling; prostate cancer; stroma